Penapisan Mikroorganisme Halofilik serta Halotoleran Penghasil Protease dari Ladang Garam Bima dengan Media Garam Krosok

Abstract

Kebutuhan protease di Indonesia semakin meningkat seiring berjalannya waktu, namun masih bergantung pada impor. Keperluan akan protease tahan garam untuk memenuhi industri tersebut, bisa didapatkan dari mikroorganisme ladang garam. Penelitian ini bertujuan mengisolasi dan mengidentifikasi mikroorganisme halofilik dan halotoleran yang diisolasi dari Ladang Garam di daerah Bima menggunakan media artifisial dengan penambahan garam krosok dari Bima untuk melihat potensi mikroorganisme tersebut sebagai penghasil enzim protease. Metode identifikasi mikroorganisme meliputi isolasi, seleksi isolat proteolitik dengan nilai IP>1, pewarnaan Gram, ekstraksi DNA, amplifikasi, dan sekuensing gen 16S rRNA. Hasil penelitian menunjukkan sebanyak 116 isolat halofilik dan 43 isolat halotoleran berhasil diisolasi pada media dengan penambahan 15%, 20%, dan 5% garam krosok Bima. Sebanyak 28 isolat bersifat proteolitik yang ditandai dengan zona bening pada media susu skim 2%. Identifikasi 8 dari 9 isolat proteolitik terpilih berdasarkan gen 16S rRNA memiliki kemiripan 99%-100%. Isolat BM3-4a, BM4-7a, BM4-2a, BM16-2a, BM4-3a, BM4-4a, dan BM7-2b mirip Halobacillus sp. dan isolat BM4-1a mirip Bacillus sp.

The demand for protease in Indonesia is growing steadily, but it was still dependent on imports. The need for salt-resistant protease to fulfill the industry could be obtained from solar salt pond microorganisms. This study aimed to isolate and identify halophilic and halotolerant microorganisms isolated from Salt Ponds in Bima using artificial media with the addition of solar salt from Bima to assess the potential of these microorganisms as protease enzyme producers. Microorganism identification methods included isolation, selection of proteolytic isolates with IP>1 value, Gram staining, DNA extraction, amplification, and 16S rRNA gene sequencing. The results showed that 116 halophilic isolates and 43 halotolerant isolates were successfully isolated on media with the addition of 15%, 20%, and 5% Bima solar salt. A total of 28 isolates were proteolytic characterized by a clear zone on 2% skim milk media. Identification of 8 out of 9 selected proteolytic isolates based on 16S rRNA gene had 99%-100% similarity. Isolates BM3-4a, BM4-7a, BM4-2a, BM16-2a, BM4-3a, BM4-4a, and BM7-2b were similar to Halobacillus sp., and isolate BM4-1a was similar to Bacillus sp.